Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H

Y. Inaguma, Y. Akahori, Y. Murayama, K. Shiraishi, S. Tsuzuki-Iba, A. Endoh, J. Tsujikawa, A. Demachi-Okamura, K. Hiramatsu, H. Saji, Y. Yamamoto, N. Yamamoto, Y. Nishimura, T. Takahashi, K. Kuzushima, N. Emi, Y. Akatsuka

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Abstract

The genetic transfer of T-cell receptors (TCRs) directed toward target antigens into T lymphocytes has been used to generate antitumor T cells efficiently without the need for the in vitro induction and expansion of T cells with cognate specificity. Alternatively, T cells have been gene-modified with a TCR-like antibody or chimeric antigen receptor (CAR). We show that immunization of HLA-A2 transgenic mice with tetramerized recombinant HLA-A2 incorporating HA-1 H minor histocompatibility antigen (mHag) peptides and β2- microglobulin (HA-1 H/HLA-A2) generate highly specific antibodies. One single-chain variable region moiety (scFv) antibody, #131, demonstrated high affinity (K D =14.9 nM) for the HA-1 H/HLA-A2 complex. Primary human T cells transduced with #131 scFV coupled to CD28 transmembrane and CD3ζ domains were stained with HA-1 H/HLA-A2 tetramers slightly more intensely than a cytotoxic T lymphocyte (CTL) clone specific for endogenously HLA-A2-and HA-1 H-positive cells. Although #131 scFv CAR-T cells required >100-fold higher antigen density to exert cytotoxicity compared with the cognate CTL clone, they could produce inflammatory cytokines against cells expressing HLA-A2 and HA-1 H transgenes. These data implicate that T cells with high-affinity antigen receptors reduce the ability to lyse targets with low-density peptide/MHC complexes (∼100 per cell), while they could respond at cytokine production level.

Original languageEnglish
Pages (from-to)575-584
Number of pages10
JournalGene therapy
Volume21
Issue number6
DOIs
Publication statusPublished - 06-2014

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All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Inaguma, Y., Akahori, Y., Murayama, Y., Shiraishi, K., Tsuzuki-Iba, S., Endoh, A., Tsujikawa, J., Demachi-Okamura, A., Hiramatsu, K., Saji, H., Yamamoto, Y., Yamamoto, N., Nishimura, Y., Takahashi, T., Kuzushima, K., Emi, N., & Akatsuka, Y. (2014). Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H. Gene therapy, 21(6), 575-584. https://doi.org/10.1038/gt.2014.30